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2026: JIPK VOLUME 18 ISSUE 1 YEAR 2026 (FEBRUARY 2026, ISSUE IN PROGRESS)

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Fisheries Biotechnology Fish Diseases Fish Health Management

Polyvalent formalin-killed Aeromonas hydrophila vaccine with oil adjuvant drives coordinated humoral, innate, and cytokine responses in giant gourami (Osphronemus goramy)

Rozi
Doctoral Program of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya 60115, Indonesia 
https://orcid.org/0000-0003-4117-1335
Wiwiek Tyasningsih
Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0003-0604-8534
Jola Rahmahani
Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0001-9103-3200
Eduardus Bimo Aksono
Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0001-6961-2556
Muchammad Yunus
Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0001-7516-6628
Mohammad Anam Al-Arif
Division of Farm, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0002-5220-9311
Suryo Kuncorojakti
Division of Veterinary Anatomy, Faculty of Veterinary Medicine, Airlangga University 60115, Surabaya, Indonesia
https://orcid.org/0000-0001-5170-9982
Daruti Dinda Nindarwi
Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0003-3353-4415
Putri Desi Wulan Sari
Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0001-8832-113X
Nina Nurmalia Dewi
Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0001-5300-7538
Woro Hastuti Satyantini
Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0002-9899-9611
Muhammad Browijoyo Santanumurti
Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0003-0846-9447
Dita Wisudyawati
Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0009-0002-4824-5931
Mohammad Noor Amal Azmai
Department of Biology, Faculty of Science, Universiti Putra Malaysia, Serdang, Malaysia
https://orcid.org/0000-0002-7664-9821
Annas Salleh
Department of Veterinary Laboratory Diagnosis, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Malaysia
https://orcid.org/0000-0003-4142-0418
Gazali Salim
Study Program of Aquatic Resources Management, Faculty of Fisheries and Marine Science, Universitas Borneo Tarakan
https://orcid.org/0000-0002-2416-905X
Suwarno Suwarno
Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia
https://orcid.org/0000-0002-4997-5242

Corresponding Author(s) : Suwarno Suwarno

suwarno@fkh.unair.ac.id

Jurnal Ilmiah Perikanan dan Kelautan, 2026: JIPK VOLUME 18 ISSUE 1 YEAR 2026 (FEBRUARY 2026, ISSUE IN PROGRESS)

Abstract

Graphical Abstract 



Highlight Research



  1. Adjuvanted polyvalent FKC elicited the strongest and most sustained multi-arm immune response in Osphronemus goramy compared with monovalent and non-adjuvanted vaccines.

  2. The lead formulation combined high agglutinating antibody titres with enhanced NBT respiratory burst, indicating synergistic humoral–innate activation against Aeromonas hydrophila.

  3. Polyvalent vaccines did not dilute immunogenicity; instead, strain combination plus adjuvant broadened and amplified immune responsiveness.

  4. Longitudinal profiling of il-1β and ifn-γ revealed a stable pro-inflammatory/Th1-like cytokine signature uniquely associated with the adjuvanted polyvalent FKC.

  5. The integrated immunological “fingerprint” supports the adjuvanted polyvalent FKC as a rational lead candidate for motile Aeromonas septicaemia control in warm-water gourami aquaculture.


 


Abstract


Bacterial septicaemia caused by Aeromonas hydrophila is an important constraint for giant gourami (Osphronemus goramy) culture in Southeast Asia. Inactivated whole-cell (bacterin) vaccines are widely used against bacterial diseases in aquaculture, but comparative data on monovalent versus polyvalent A. hydrophila vaccines, with and without oil-based adjuvant, remain scarce for this species. This study evaluated the safety, immunological responses, and protective efficacy of three formalin-killed cell (FKC) vaccines prepared from gourami-derived A. hydrophila isolates: a monovalent FKC (P2), a non-adjuvanted polyvalent FKC (P3), and an oil-adjuvanted polyvalent FKC (P4), using PBS (P1) as a control. Sub-adult giant gourami were vaccinated intraperitoneally and monitored for 42 days. Serum agglutinating antibody titres, nitroblue tetrazolium (NBT)-reducing activity, and splenic il-1β and ifn-γ mRNA expression were measured at multiple time points. At 21 days post-vaccination, a separate cohort was challenged intraperitoneally with virulent A. hydrophila Ah-S1, and survival was recorded for 14 days; relative percent survival (RPS) was calculated at day 14. All FKC formulations were clinically well tolerated, with only transient post-vaccination inappetence and no gross injection-site pathology. Vaccination induced clear, treatment- and time-dependent increases in agglutinating antibody titres, NBT-reducing activity, and splenic il-1β and ifn-γ transcription, with the strongest and most sustained responses in P4, intermediate responses in P3 and P2, and minimal changes in P1. Following homologous challenge, day-14 survival was 8.3% in P1, 61.7% in P2, 75.0% in P3, and 83.3% in P4, with corresponding RPS values of 58.2%, 72.7%, and 81.8%, respectively. Under these experimental conditions, the oil-adjuvanted polyvalent FKC (P4) produced the greatest enhancement of immune responses and protection against intraperitoneal A. hydrophila challenge in giant gourami. These findings support this formulation as a candidate for further vaccine development in O. goramy and highlight the need for dose optimisation, safety assessment, heterologous challenge, and field-validation studies before recommendations for large-scale use in aquaculture.

Keywords

Aeromonas hydrophila vaccine Adjuvanted polyvalent FKC
Rozi, Wiwiek Tyasningsih, Jola Rahmahani, Eduardus Bimo Aksono, Muchammad Yunus, Mohammad Anam Al-Arif, Suryo Kuncorojakti, Daruti Dinda Nindarwi, Putri Desi Wulan Sari, Nina Nurmalia Dewi, Woro Hastuti Satyantini, Muhammad Browijoyo Santanumurti, Dita Wisudyawati, Mohammad Noor Amal Azmai, Annas Salleh, Gazali Salim, & Suwarno, S. (2025). Polyvalent formalin-killed Aeromonas hydrophila vaccine with oil adjuvant drives coordinated humoral, innate, and cytokine responses in giant gourami (Osphronemus goramy). Jurnal Ilmiah Perikanan Dan Kelautan. Retrieved from https://e-journal.unair.ac.id/JIPK/article/view/82866
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Author Biographies

Rozi, Doctoral Program of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya 60115, Indonesia 

Doctoral Program of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya 60115, Indonesia 

Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Wiwiek Tyasningsih, Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Jola Rahmahani, Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Eduardus Bimo Aksono, Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Muchammad Yunus , Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Mohammad Anam Al-Arif, Division of Farm, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Division of Farm, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Suryo Kuncorojakti, Division of Veterinary Anatomy, Faculty of Veterinary Medicine, Airlangga University 60115, Surabaya, Indonesia

Division of Veterinary Anatomy, Faculty of Veterinary Medicine, Airlangga University 60115, Surabaya, Indonesia

Daruti Dinda Nindarwi, Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Putri Desi Wulan Sari, Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Nina Nurmalia Dewi, Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Woro Hastuti Satyantini, Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Muhammad Browijoyo Santanumurti, Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Dita Wisudyawati, Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Department of Aquaculture, Faculty of Fisheries and Marine, Airlangga University, Surabaya 60115, Indonesia

Mohammad Noor Amal Azmai, Department of Biology, Faculty of Science, Universiti Putra Malaysia, Serdang, Malaysia

Department of Biology, Faculty of Science, Universiti Putra Malaysia, Serdang, Malaysia

Annas Salleh, Department of Veterinary Laboratory Diagnosis, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Malaysia

Department of Veterinary Laboratory Diagnosis, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Malaysia

Gazali Salim, Study Program of Aquatic Resources Management, Faculty of Fisheries and Marine Science, Universitas Borneo Tarakan

Study Program of Aquatic Resources Management, Faculty of Fisheries and Marine Science, Universitas Borneo Tarakan

Suwarno Suwarno, Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

Division of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia

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